ICP22 of Herpes Simplex Virus 1 Decreases RNA Polymerase Processivity.

To determine the role of ICP22 in transcription, we performed precise nuclear run-on followed by deep sequencing (PRO-seq) and global nuclear run-on with sequencing (GRO-seq) in cells infected with a viral mutant lacking the entire ICP22-encoding α22 (US1/US1.5) gene and a virus derived from this mutant bearing a restored α22 gene. At 3 h postinfection (hpi), the lack of ICP22 reduced RNA polymerase (Pol) promoter proximal pausing (PPP) on the immediate early α4, α0, and α27 genes. Diminished PPP at these sites accompanied increased Pol processivity across the entire herpes simplex virus 1 (HSV-1) genome in GRO-seq assays, resulting in substantial increases in antisense and intergenic transcription. The diminished PPP on α gene promoters at 3 hpi was distinguishable from effects caused by treatment with a viral DNA polymerase inhibitor at this time. The ICP22 mutant had multiple defects at 6 hpi, including lower viral DNA replication, reduced Pol activity on viral genes, and increased Pol activity on cellular genes. The lack of ICP22 also increased PPP release from most cellular genes, while a minority of cellular genes exhibited decreased PPP release. Taken together, these data indicate that ICP22 acts to negatively regulate transcriptional elongation on viral genes in part to limit antisense and intergenic transcription on the highly compact viral genome. This regulatory function directly or indirectly helps to retain Pol activity on the viral genome later in infection. IMPORTANCE The longstanding observation that ICP22 reduces RNA polymerase II (Pol II) serine 2 phosphorylation, which initiates transcriptional elongation, is puzzling because this phosphorylation is essential for viral replication. The current study helps explain this apparent paradox because it demonstrates significant advantages in negatively regulating transcriptional elongation, including the reduction of antisense and intergenic transcription. Delays in elongation would be expected to facilitate the ordered assembly and functions of transcriptional initiation, elongation, and termination complexes. Such limiting functions are likely to be important in herpesvirus genomes that are otherwise highly transcriptionally active and compact, comprising mostly short, intronless genes near neighboring genes of opposite sense and containing numerous 3'-nested sets of genes that share transcriptional termination signals but differ at transcriptional start sites on the same template strand.

Immediate Early Proteins of Herpes Simplex Virus Transiently Repress Viral Transcription before Subsequent Activation.

Herpes simplex virus 1 (HSV-1) utilizes cellular RNA polymerase II (Pol) to transcribe its genes in one of two phases. In the latent phase, viral transcription is highly restricted, but during the productive lytic phase, more than 80 genes are expressed in a temporally coordinated cascade. In this study, we used Precision nuclear Run On followed by deep Sequencing (PRO-Seq) to characterize early viral transcriptional events using HSV-1 immediate early (IE) gene mutants, corresponding genetically repaired viruses, and wild-type virus. Unexpectedly, in the absence of the IE genes ICP4, ICP22, and ICP0 at 1.5 hours postinfection (hpi), we observed high levels of aberrant transcriptional activity across the mutant viral genomes but substantially less on either wild-type or the congenic repaired virus genomes. This feature was particularly prominent in the absence of ICP4 expression. Cycloheximide treatment during infection with both the ICP4 and ICP22 mutants and their respective genetic repairs did not alter the relative distribution of Pol activity, but it increased overall activity across both viral genomes, indicating that both virion components and at least some de novo protein synthesis were required for full repression. Overall, these data reveal that prior to their role in transcriptional activation, IE gene products and virion components first repress transcription and that the HSV-1 lytic transcriptional cascade is mediated through subsequent derepression steps. IMPORTANCE HSV-1 transcription during productive replication is believed to comprise a series of activation steps leading to a specific sequence of gene expression. Here, we show that virion components and IE gene products ICP0, ICP4, and ICP22 first repress viral gene transcription to various degrees before subsequently activating specific gene subsets. It follows that the entire HSV transcriptional program involves a series of steps to sequentially reverse this repression. This previously uncharacterized repressive activity of IE genes very early in infection may represent an important checkpoint allowing HSV-1 to orchestrate either the robust lytic transcriptional cascade or the more restricted transcriptional program during latency.

Canadian pediatric eating disorder programs and virtual care during the COVID-19 pandemic: a mixed-methods approach to understanding clinicians' perspectives.

BACKGROUND: As a result of the public health measures put in place during the COVID-19 pandemic in Canada, many health services, including those for the treatment of eating disorders, were provided at a distance. This study aims to describe the adaptations made in specialized pediatric eating disorder programs in Canada and the impact of these adaptations on health professionals' experience of providing care. METHODS: A mixed-methods design was used to survey healthcare professionals working in specialized pediatric eating disorder programs about adaptations to treatment made during the pandemic and the impact of these adaptations on their experience of providing care. Data were collected between October 2021 and March 2022 using a cross-sectional survey comprising 25 questions and via semi-structured interviews. Quantitative data were summarized using descriptive statistics and qualitative data were interpreted using qualitative content analysis. RESULTS: Eighteen healthcare professionals in Canada completed the online survey, of whom six also participated in the semi-structured interviews. The cross-sectional survey confirmed that, unlike in pre-pandemic times, the majority of participants provided medical care (15/18) and mental health care (17/18) at a distance during the pandemic, with most participants using telephone (17/18) and videoconferencing (17/18). Most (16/18) health professionals indicated that virtual care would continue to be used as a tool in pediatric ED treatment after the pandemic. Participants used a combination of virtual and in-person care, with most reporting weighing patients both in clinic (16/18) and virtually (15/18). Qualitative content analysis generated five themes: (1) responding to increased demand with insufficient resources; (2) adapting to changes in care due to the COVID-19 pandemic; (3) dealing with uncertainty and apprehension; (4) virtual care as an acceptable and useful clinical tool, and (5) optimal conditions and future expectations. Most interview participants (5/6) had globally positive views of virtual care. CONCLUSIONS: Providing virtual multidisciplinary treatment for children and adolescents with eating disorders seemed feasible and acceptable to professionals during the pandemic. Moving forward, focusing on health professionals' perspectives and providing appropriate training in virtual interventions is essential given their central role in successful implementation and continued use of virtual and hybrid care models.

Childhood hyperactivity, eating behaviours, and executive functions: Their association with the development of eating-disorder symptoms in adolescence.

BACKGROUND: Cross-sectional studies have shown that hyperactivity and impaired executive functioning are associated with symptoms of eating disorders in adolescence and adulthood. Whether hyperactivity and executive functions in early life can prospectively predict the emergence of eating disorder symptoms in adolescence remains unknown. The present study relies on a longitudinal design to investigate how hyperactivity at age 3, eating behaviours at age 3.5 and cognition at ages 3-6 were associated with the development of eating-disorder symptoms from 12 to 20 years old. METHODS: Using archival data collected since 1997 from the Quebec Longitudinal Study of Child Development cohort (N = 2, 223), we used Latent Curve Models to analyse predictors of youth's trajectories of eating-disorder symptoms at four timepoints. RESULTS: A quadratic (curvilinear) trajectory of eating-disorder symptoms was found to be most representative of the data. Higher hyperactivity at age 3 was associated with higher levels of eating-disorder symptoms at age 12, and this association was partially mediated by higher levels of overeating and cognitive inflexibility in childhood. Cognitive inflexibility in childhood also mediated the association between hyperactivity at age 3 and increases in eating-disorder symptoms during adolescence. Furthermore, working memory was indirectly related to eating-disorder symptoms via the mediational role of cognitive flexibility. CONCLUSIONS: Hyperactivity, overeating, cognitive inflexibility, and working memory early in life might precede the onset of eating-disorder symptoms in adolescence. Early behavioural and cognitive screening may help to identify children who are most at risk for eating disorders. This, in turn, could guide preventive interventions.

Eating-disorder symptoms, such as body image issues, maladaptive behaviors, and preoccupation with weight, tend to develop in adolescence. However, it is unclear whether early childhood characteristics or behaviours could be indicators of a risk of developing eating-disorder symptoms later. The current study examined the possible link between certain early behaviours (e.g., hyperactivity, childhood eating), early cognitive processes, and eating-disorder symptoms development in a community cohort followed from birth. Results showed that being hyperactive in early childhood predicts higher levels of eating-disorder symptoms at the beginning of adolescence (age 15), and that this is partially explained by a link between being hyperactive, being more rigid in our ways of thinking, and engaging in overeating behaviours. Additionally, more early rigid ways of thinking predicted the increase in symptoms over time. Our results demonstrate possible behaviours and characteristics that could be used to identify children at risk of eating disorders, which in future research could potentially help improve our preventive interventions.

The use of technology in the treatment of youth with eating disorders: A scoping review.

BACKGROUND: Adolescence and young adulthood is a high-risk period for the development of eating disorders. In recent years, there has been an increase in use of technology-based interventions (TBIs) for the treatment of eating disorders. The objective of this study was to determine the types of technology used for eating disorder treatment in youth and their effectiveness. METHODS: A scoping review was conducted according to PRISMA-ScR guidelines. Four databases were searched. Eligible articles included: (1) a TBI (2) participants with a mean age between 10- and 25-years and meeting DSM-IV or DSM-5 criteria for any eating disorder and (3) qualitative or quantitative designs. Quantitative and qualitative studies were assessed for quality. RESULTS: The search identified 1621 articles. After screening of titles and abstracts, 130 articles were read in full and assessed for eligibility by two raters. Forty-nine (29 quantitative and 20 qualitative, observational, or mixed methods studies) met inclusion criteria. Quality ratings indicated that 78% of quantitative studies had a low risk of bias and 22% had a moderate risk. Technologies reviewed in our study included videoconference therapy, mobile applications, and online self-help. We considered interventions used both within sessions with clinicians as well as those used in between sessions by patients alone. Fifteen of 18 (83%) quantitative studies found that TBIs reduce eating disorder symptomatology, with nine of those reporting medium-to-large effect sizes. Qualitative data was of high quality and suggested that virtual interventions are acceptable in this population. CONCLUSIONS: Although identified studies are of high quality, they are limited in number. More research is needed, particularly regarding videoconferencing and mobile applications. Nonetheless, TBIs show promise for the treatment of eating disorders in youth. TRIAL REGISTRATION: Not applicable.

The use of virtual care to treat adolescents and young adults with eating disorders has increased in recent years. To better understand what types of technologies are being used in virtual care and these technologies' effectiveness, we systematically reviewed research studies that discussed the use of any technology to treat youth (which was defined as young people aged 25 years or under) with an eating disorder. We found 49 relevant studies. They revealed that videoconference therapy, mobile applications, and online self-help are most frequently used for virtual treatment. Of the 18 studies evaluating the effectiveness of these technologies, 15 found virtual care to be effective for reducing eating disorder symptoms. Studies describing patient experiences with these technologies, of which there were nine, suggested that patients overall were satisfied with receiving virtual care. Although these results are promising, research on virtual treatment is still lacking, especially on videoconferencing therapy and mobile applications.

An Intensive Ambulatory Care Program for Adolescents With Eating Disorders Combining In-Person and Web-Based Care: Protocol for a Single-Site Naturalistic Trial.

BACKGROUND: The incidence of eating disorders (EDs) among adolescents has significantly increased since the beginning of the COVID-19 pandemic. Hybrid care, which combines web-based and in-person modalities, is a promising approach for adolescents with EDs but remains understudied in this population. OBJECTIVE: We aimed to implement a novel hybrid (web-based and in-person) intensive ambulatory care program for youth and evaluate its feasibility, acceptability, and preliminary effectiveness. METHODS: We will use a naturalistic pretest-posttest design to evaluate our proposed pilot Intensive Ambulatory Care Program (IACP). This novel type of day hospital care follows evidence-based principles and uses a family-centered, educational, and motivational approach. It will be tailored to the psychological needs of each participant and will be delivered in a hybrid format. A total of 100 participants meeting the DSM-5 (Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition) criteria for EDs, aged 12-18 years, will be recruited over the 2-year trial period. We will examine recruitment, retention, and adhesion-to-protocol rates; participant and family satisfaction; and preliminary effectiveness using quantitative self-report questionnaires. RESULTS: Rolling recruitment will take place from winter 2022 to fall 2023, during which time we expect to recruit approximately 80% (100/120) of eligible participants, retain at least 75% (75/100) of enrolled participants and have at least 70% (70/100) of enrolled participants complete at least one therapeutic session per week and all pre- and postintervention questionnaires. Data collection will occur concurrently. We base our recruitment and retention estimates on previous literature and consider that the highly flexible design of the IACP and the fact that no extra work will be required of individuals in the program to participate in the study, will lead to high levels of feasibility. We anticipate that participants and their families will be satisfied with both the program and hybrid delivery format. We expect that participation in the IACP will be associated with a medium effect size reduction in ED psychopathology from baseline to end of treatment. The data analysis and manuscript writing are expected to be completed by the summer of 2024. CONCLUSIONS: Given the high clinical burden associated with EDs, this study has the potential to fill an important research gap by testing the implementation of a novel hybrid mode of intervention. If feasible, acceptable, and effective, the IACP could lead to important improvements in health care services for adolescents with EDs. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): PRR1-10.2196/37420.

Developmental trajectories of eating disorder symptoms: A longitudinal study from early adolescence to young adulthood.

BACKGROUND: Adolescence is a critical period for the development of eating disorders, but data is lacking on the heterogeneity of their evolution during that time-period. Group-based trajectories can be used to understand how eating disorders emerge and evolve over time. The aim of this study was to identify groups of individuals with distinct levels of eating disorder symptoms between 12 and 20 years and the onset of different types of symptoms. We also studied sex differences in the evolution and course of eating disorder symptoms from early adolescence to adulthood. METHODS: Using archival data from the QLSCD cohort, trajectories of eating disorder symptomatology were estimated from ages 12 to 20 years using semiparametric models. These trajectories included overall eating disorder symptomatology as measured by the SCOFF (Sick, Control, One Stone, Fat, Food), sex, and symptom-specific trajectories. RESULTS: Two groups of adolescents following distinct trajectories of eating disorder symptoms were identified. The first trajectory group included 30.9% of youth with sharply rising levels between 12 and 15 years, followed by high levels of symptoms between 15 and 20 years. The second trajectory group included 69.1% of youth with low and stable levels of symptoms between 12 and 20 years. Sex-specific models indicated that the proportion of girls in the high trajectory group was 1.3 times higher than the proportion of boys (42.8% girls vs. 32.3% boys). Trajectories of SCOFF items were similar for loss-of-control eating, feeling overweight, and attributing importance to food. The weight loss item had a different developmental pattern, increasing between 12 and 15 years and then decreasing between 17 and 20 years. CONCLUSIONS: The largest increase in eating disorder symptoms in adolescence is between the ages of 12 and 15 . Yet, most prevention programs start after 15 years of age. Our findings suggest that, unlike common practices, eating disorder prevention programs should aim to start before puberty.

Eating disorders, typically involving preoccupation with eating, excessive exercise, and body image issues, are particularly common in adolescence. However, their evolution over time remains unclear as certain signs and symptoms may appear sooner than others. The current study studied the development of eating disorder symptoms in a community cohort followed from birth to adulthood. This study describes trajectories of eating disorder symptoms and risk from age 12 to 20 in both boys and girls. Results showed that the largest increase in eating disorder symptoms occurs between 12 and 15 years of age, both in overall symptomatology as well as in specific symptoms such as loss-of-control when eating, feeling overweight, and attributing importance to food. Additionally, more girls than boys appeared at risk for eating disorders, although the patterns represented by the trajectories were similar in both sexes. Our results highlight the importance of starting early prevention programs before the beginning of adolescence, when symptoms usually start to manifest.